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Background and Objective: Inhibitor of differentiation or DNA binding -1 (Id-1) has been shown to be increased in several types of advanced cancer, and to be associated with aggressive and metastatic abilities of cancer cells. Recently, more and more evidence indicates that epithelial-to-mesenchymal transition (EMT) is an important mechanism taking place during tumor invasion and metastasis, but the molecular pathways underlying EMT have not been clearly established. This study was to investigate the expression of Id-1 in bladder cancer and its association with EMT., Materials and Methods: A total of 169 tissues, consisting of 147 primary bladder cancers and 22 adjacent normal tissues were included in this study. Id-1, E-cadherin, and β-catenin were examined immunohistochemically in paraffin sections. The pBabe-Id-1 expression retroviral vector and retroviral vectors containing an Id-1-specific small interfering RNA oligonucleotides (si-Id-1) were transfected into 2 bladder cancer cell lines respectively. Then, we used Western blotting and immunofluorescent staining to detect the cellular expression of epithelial markers and mesenchymal markers. The invasion and migration ability of bladder cancer cells were identified by type I collagen invasion assay and wound closure assay., Results: We demonstrated that increased Id-1 expression was associated with advanced tumor stage and grade. In addition, the increased Id-1 expression in bladder tumors was also correlated with decreased membranous E-cadherin and β-catenin expression. In vitro, studies showed that inactivation of the Id-1 gene conferred morphologic transition of bladder cancer cells from a fibroblastic to epithelial appearance, and overexpression of Id-1 could lead to acquisition of a fibroblastic spindle cell phenotype accompanied by loss of cell-to-cell contacts. By Western blotting and immunofluorescent staining, we showed that the expression level of Id-1 was correlated with the expression of mesenchymal markers but was inversely correlated with the expression of epithelial markers. Moreover, results of collagen invasion and wound closure assays showed ectopic Id-1 expression led to increased ability of invasion and migration., Conclusions: Our results suggest that Id-1 may play roles in tumor progression and EMT activation in bladder cancer., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Growth curve of bacteria treated by the rare earth elements and calmodulin inhibitor (chlorpromazine) was studied, and the effect of REE on calf thymus-DNA and intracellular DNA in bacteria was studied by fluorescence spectrum. The results showed that the logarithmic phase of growth and fission of bacteria can be advanced by REE, so they grew faster than contrast. At the same time, Bacillus subtilis was found bigger by SEM. But growth and fission was inhibited by chlorpromazine, indicating that the mechanism about growth and fission of bacteria by REE was connected with calmodulin inhibitor. Fluorescence of ct-DNA can be enhanced by REE. But intracellular DNA in bacteria was different, it can be enhanced by Ce(NO3)3, quenched by La(NO3)3. So we think that the function of REE to biological materials was very complicated.

Objective: To observe the dynamic changes of proliferation and differentiation of neural stem cells (NSCs) in subventricular zone (SVZ) and dentate gyrus (DG) in vascular dementia (VD) rats., Methods: VD models were established by repeatedly clipping the common carotid arteries of the rat in combination with an intraperitoneal injection of sodium nitroprusside solution in anesthetized SD rats. Morris maze test was used to detect the learning and memory ability of the rats and immune fluorescence single and double labeling method to detect the proliferation and differentiation of neural stem cells and the expression of neurogranin (Ng) at 15 d, 1 month, 2 month, 4 month time points., Results: Compared with sham-operated group, the escape latency of model group rats were significantly longer at all the time points (P < 0.01). The BrdU positive cells in SVZ and DG of VD model groups were markedly increased in comparison with sham-operated group at 15 d and 1 month time point (P < 0.01), and the number of BrdU positive cells in SVZ of model groups were still larger than that of sham-operated group at 2 month and 4 month time point (P < 0.01). In model group, the number of the BrdU/Ng double staining cells were increased and higher than that in sham-operated groups (P < 0.05)., Conclusion: The proliferation of NSCs can be enhanced noticeably in a certain time in SVZ and DG region and NSCs differentiate into mature neurons with the expression of Ng in DG region in VD rats, which may play some compensatory roles in the nerve regeneration and functional repairmen after cerebral injury of VD.

In order to automatically detect hemorrhages in fundus images, and develop an automated diabetic retinopathy screening system, a novel algorithm named locally adaptive region growing based on multi-template matching was established and studied. Firstly, spectral signature of major anatomical structures in fundus was studied, so that the right channel among RGB channels could be selected for different segmentation objects. Secondly, the fundus image was preprocessed by means of HSV brightness correction and contrast limited adaptive histogram equalization (CLAHE). Then, seeds of region growing were founded out by removing optic disc and vessel from the resulting image of normalized cross-correlation (NCC) template matching on the previous preprocessed image with several templates. Finally, locally adaptive region growing segmentation was used to find out the exact contours of hemorrhages, and the automated detection of the lesions was accomplished. The approach was tested on 90 different resolution fundus images with variable color, brightness and quality. Results suggest that the approach could fast and effectively detect hemorrhages in fundus images, and it is stable and robust. As a result, the approach can meet the clinical demands.

Objective: To investigate the effect of demethylating agent 5-aza-2'-deoxycytidine (5-Aza-CdR) on the growth of human pancreatic cancer cell line MiaPaca2 and the expression and methylation of tumor suppressor gene RUNX3., Methods: Human pancreatic cancer cell line MiaPaca2 cells were treated with different concentrations of 5-Aza-CdR. Morphological changes of MiaPaca2 cells were observed by light microscopy. The activity of cell proliferation was analyzed by MTT assay. The changes of RUNX3 mRNA expression were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Changes of RUNX3 gene methylation was detected by methylation-specific polymerase chain reaction., Results: MiaPaca2 cells were treated with 2.5, 5, 10 and 20 µmo1/L 5-Aza-CdR, respectively. The inhibition rates of MiaPaca2 cells treated for 24 h were (9.17 ± 2.15)%, (10.75 ± 2.04)%, (12.57 ± 1.64)% and (18.70 ± 1.51)%, respectively. The inhibition rates were (14.94 ± 1.68)%, (18.60 ± 1.57)%, (22.84 ± 1.58)% and (33.24 ± 1.53)%, respectively, after 48 h treatment; (21.46 ± 1.60)%, (28.62 ± 1.72)%, (35.14 ± 1.64)% and (45.06 ± 1.47)%, respectively, after 72 h treatment; and (26.35 ± 1.71)%, (34.48 ± 1.69)%, (40.05 ± 1.60)% and (49.99 ± 1.61)%, respectively, after 96 h treatment. The differences between inhibition rates of each experimental and control groups (0.00 ± 0.00)% were statistically significant (P < 0.05). At the same time, the inhibition rates of different concentration groups showed significant differences (P < 0.05). At 48 h, 72 h and 96 h, the inhibition rates of each pair concentration groups showed significant differences (P < 0.05). 5-Aza- CdR inhibited the growth of MiaPaca2 cells, and the higher the concentration, the stronger the inhibition after 24 h. 5-Aza-CdR also reversed the methylation status of RUNX3 gene, and restored the expression of RUNX3 mRNA with a dose-effect relationship., Conclusions: The methylation of RUNX3 gene is significantly related with the occurrence and development of pancreatic cancer, and abnormal methylation of RUNX3 gene may contribute to the loss of RUNX3 mRNA expression. 5-Aza-CdR may effectively cause reversion of RUNX3 methylation, and treatment with 5-Aza-CdR can reactivate the gene expression and inhibit the cell growth. This may provide a new way for diagnosis and treatment of pancreatic cancer.

Background: Acute rejection remains an important cause of renal allograft dysfunction and the need for accurate diagnosis is essential to successfully treat transplant recipients. The purpose of this study was to determine the costimulatory molecules OX40 and OX40L messenger RNA (mRNA) levels in peripheral blood mononuclear cells (PBMCs) to predict acute renal transplant rejection., Methods: The whole blood samples from 20 recipients with biopsy-confirmed acute rejection (rejection group), 20 recipients with stable graft function and normal biopsy results (stable group) after kidney transplantation, and 20 healthy volunteers (control group) were collected. The mRNA levels of OX40 and OX40L were analyzed with TaqMan real-time reverse transcriptase polymerase chain reaction (RT-PCR). The association of OX40 and OX40L mRNA levels with disease severity was investigated., Results: There was no significant difference of OX40, OX40L mRNA levels in PBMCs between the stable group and control group (P > 0.05). The levels of OX40 and OX40L mRNA were significantly higher in the rejection group than in the control group (P < 0.01 and P < 0.05, respectively). Non-significantly higher OX40L mRNA and significantly higher OX40 mRNA in PBMCs were observed in subjects in the rejection group compared with the stable group (P > 0.05 and P < 0.01, respectively). Receiver operating characteristic (ROC) curve analysis demonstrated that OX40 mRNA levels could discriminate recipients who subsequently suffered acute allograft rejection (area under the curve, 0.908). OX40 and OX40L mRNA levels did not significantly correlate with serum creatinine levels in the rejection group (P > 0.05). Levels of OX40 mRNA after anti-rejection therapy were lower than those at the time of protocol biopsy in the rejection group (P < 0.05)., Conclusion: Our data suggest that measurement of OX40 mRNA levels after transplant might offer a noninvasive means for recognizing recipients at risk of acute renal allograft rejection.

The technology of automated detection of diabetic retinopathy (DR) on fundus retinal images can not only make mass screening possible, but also offer a powerful adjunct for early diagnosis, treatment of diabetic retinopathy, and scientific research on human vision. For this purpose, an algorithm based on mathematical morphology for automated detection of diabetic retinopathy was proposed. Firstly, the optic disc was segmented by mathematical morphology and threshold in order to find candidate regions possibly containing lesions. Secondly, some methods such as morphological reconstruction were applied to find the exact contours of lesions. Finally, the true lesions were found out exactly. Experimental results showed that the algorithm was fast and effective in detecting diabetic retinopathy of fundus retinal images.

Studies have shown that the expression of inhibitor of differentiation (Id-1) is increased in bladder cancer and is associated with drug resistance. S-allylmercaptocysteine (SAMC), a water-soluble component of garlic, is known to have a potent therapeutic effect on human cancer. The aim of this study was to investigate whether Id-1 expression mediates SAMC-induced cell death in bladder cancer cells. After generating stable Id-1-expressing and si-Id-1 transfectants in various bladder cancer cell lines, cell sensitivity to SAMC was compared by colony formation and MTT assays. The results indicated that Id-1 overexpression reduced the positive effect of SAMC on cell survival, while the inactivation of Id-1 increased cellular susceptibility to SAMC. Using DAPI staining, the apoptosis of bladder cancer cells induced by SAMC was shown to be negatively regulated by Id-1 expression. The expression of apoptosis-related proteins analyzed by Western blotting further supported the negative role of Id-1 in SAMC-induced apoptosis. Furthermore, by wound closure and type I collagen invasion assays, the inhibitory effect of SAMC on the invasion and migration of bladder cancer cells was found to be associated with the down-regulation of Id-1. Our results demonstrated that SAMC-induced apoptosis is associated with the Id-1 pathway, and that the inactivation of Id-1 enhances the ability of SAMC to inhibit the survival, invasion and migration of bladder cancer cells. These findings may lead to the development of novel therapeutic strategies for the treatment of bladder cancer.

Aim: To investigate the mechanism of proliferation effect induced by (R,R)-XY-10 and (S,S)-XY-10 on retinal pigmented epithelial cells (ARPE-19)., Methods: Human retinal pigmented epithelial cells (ARPE-19) and human umbilical vein endothelial cells (HUVECs) were used to investigate the effect of (R,R)-XY-10 and (S,S)-XY-10 on cell growth, and their mechanisms of proliferative action by using ERK, AKT, PI3K, Protein kinase C (PKC) and Nitric oxide synthase (NOS) inhibitors., Results: (R,R)-XY-10 and (S,S)-XY-10 dose-dependently increased ARPE-19 cell proliferation, but not on HUVECs. When treated with proliferative inhibitors, H-7 (5µmol/L), hypericin (20µmol/L), PD98059 (2µmol/L), LY294002 (50µmol/L), SH-5 (10µmol/L) and L-NAME (100µmol/L), the proliferative effect was reduced by H-7, hypericin, PD98059 and LY294002, but not by SH-5 and L-NAME., Conclusion: (R,R)-XY-10 and (S,S)-XY-10 can induce cell proliferation through MAPK and PI3K dependent pathway.

Objective: To investigate the clinical features and detection of pantothenate kinase 2 (PANK2) gene mutation in a Chinese patient with Hallervorden-Spatz syndrome (HSS)., Methods: The clinical features were analyzed in one HSS patient. PANK2 gene mutations were detected by polymerase chain reaction (PCR) and DNA sequence analysis in this patient, her parents and 50 unrelated healthy persons., Results: The main symptoms of this patient were involuntary movements, dysarthria and progressive course. MRI scans showed hypointensity with a central region of hyperintensity in medial globus pallidus on T2 and T2-weighted fluid attenuated inversion recovery (FLAIR) images, i.e. "eye-of-the-tiger" sign. Novel compound heterozygous PANK2 gene mutations, G115T and A803G, were found in this patient, leading to substitution of a glutamic acid for a premature stop codon at amino acid 39 (E39X) and an aspartic acid for glycine codon at amino acid 268 (D268G) respectively. The father was a heterozygote for G115T mutation and the mother a heterozygote for A803G mutation., Conclusion: PANK2 gene mutations are present in Chinese HSS patients. And A803G mutation of PANK2 gene is probably a hot spot.

Objective: To objectively assess the effect of Qiming Granule in the treatment of diabetic retinopathy (DR) by fluorescence fundus angiography (FFA)., Methods: In a multi-center, randomized, parallel controlled clinical trial, patients with DR were randomly assigned to the control group (calcium dobesilate capsule) and the test group (Qiming Granule). Changes in the retinal blood circulation time were recorded by FFA after 3 months of medication., Results: Significant reduction was observed in the retinal arterio-venous circulation time (AVCT) in both groups (P<0.01), the value was 7.635+/-3.149 s before treatment and 5.165 +/-3.382 s after treatment in the treated group, and 7.737+/-3.413 s and 5.313+/-3.472 s in the control group respectively. Qiming Granule also reduced the arm-to-retinal circulation time (ARCT, P<0.05). The value was 17.867+/-3.872 s before treatment and 15.643+/-4.648 s after treatment in the treated group, and 17.217+/-3.833 s and 16.312+/-3.613 s in the control group (P>0.05) respectively. The ARCT in the tested group was reduced, with a statistically significant difference post-medication (P<0.01)., Conclusion: As a Chinese medicine complex prescription, Qiming Granule may alleviate retinal hypoxia and ischemia by increasing retinal blood flow and improving the blood circulation.

Recurrence and progression are the major problems in the treatment of bladder cancer. Increased expression of Id-1, a basic helix-loop-helix transcription factor, has recently been shown in several types of advanced cancer. Some studies have provided evidence to suggest that Id-1 can be considered a potential therapeutic target. The objective of this study was to investigate the role of Id-1 in the chemosensitivity of bladder cancer cells, and the effect of Id-1 on chemotherapeutic drug-induced apoptosis in bladder cancer cells. We compared the different sensitivity to epirubicin in RT112 and MGH-U1 cell lines with different Id-1 expression. Then, we transfected different vectors into RT112 and MGH-U1 respectively, and generated the stable Id-1 up-regulation and down-regulation transfectants. The results of cell viability assay showed up-regulation of Id-1 in RT112 leading to increased sensitivity in response to epirubicin, and down-regulation of Id-1 increased cellular sensitivity to epirubicin. Furthermore, the analysis of apoptosis related protein revealed that up-regulation of Id-1 suppressed epirubicin-induced apoptosis and down-regulation of Id-1 leading to increased epirubicin-induced apoptosis. Wound closure assay showed up-regulation of Id-1 leading to improved migration abilities of bladder cancer cells under chemotherapy. Our results suggest that up-regulation of Id-1 in bladder cancer cells lead to increased cell viability in response to epirubicin by its improved anti-apoptotic role, and down-regulation of Id-1 increases cellular sensitivity to epirubicin by increased anticancer drug-induced apoptosis.

Objective: To investigate one-staged orthognathic surgery and arthroscope-assisted condylar reconstruction with costochondral graft for temporomandibular diseases combined with dental-maxillofacial deformities., Methods: The preoperative measurement and model surgery was performed. The patients underwent arthroscope assisted condylar reconstruction with costochondral graft combined with bimaxillary surgery or mandibular surgery., Results: From Sept. 2003 to Dec. 2005, 15 cases were treated successfully with no bleeding, nerve injury and infection. The patients were followed up for 29 to 52 months (mean, 31.8 months). The appearance was markedly improved. TMJ function was good with no pain. The average maximum interincisal opening was 33.6 mm., Conclusion: Compared with traditional operation, one-staged orthognathic surgery and arthroscope-assisted condylar reconstruction with costochondral graft has its advantages. Indication selection and preoperative design are very important.

Objective: This is a preliminary study of the endoscopic technique on open surgery. The purpose of this study is to introduce the use of endoscopic technique for the reconstruction of the condyle with costochondral graft through a preauricular incision., Methods: 33 patients of temporomandibular joint diseases (n=49) underwent reconstruction of mandibular condyle with costochondral graft. After preparation of the recipient site through a preauricular incision, maxillo-mandibular fixation, and preparation of the costochondral graft, the graft was fixed to the lateral side of the mandible ramus under the supervision of arthroscope., Results: All the patients had successful reconstruction of the condyle with costochondral graft without any severe bleeding or craniocerebral injury., Conclusion: This case series demonstrates the feasibility of endoscopic technique for the reconstruction of the condyle through a preauricular incision. It has the advantage of high efficiency, minimal postoperative morbidity, great patient comfort, and little appearance impairment. The endoscopic technique has a promising future.

Aim: To investigate the effects of ginkgo biloba extract 761 (EGb761) on synaptic plasticity in hippocampus of vascular dementia (VD) rats., Methods: The escape latency (EL) of Morris water maze (MWM) task was measured at different time points (4 W, 8 W and 16 W), and the population spikes (PS) of granule cell layer in the dentate gyrus were induced by single pulse stimulation to perfo rate path fibers before and after high frequency stimulation (HFS) in vivo., Results: MWM test showed that the escape latency (EL) of VD model group were highly longer than that of the sham-operated group, while the EL of EGb761-treated group was significantly shorter than that of model group, but still longer than that of the sham-operated group. The incidence rates of LTP induction in 1 m, 2 m and 4 m subgroups of model group were significantly lower than that of sham-operated group and EGb761-treated group at different time point. The relative amplitudes of PS after HFS in 1 m, 2 m and 4 m subgroups of model group were obviously reduced compared with that of the corresponding subgroups of sham-operated group and EGb761-treated group. There was no obvious significance in the peak latency of PS between different subgroups and different LTP-tested time point., Conclusion: VD model rats had apparent and long-lasting dysfunction of learning and memory, EGb761 could accelerate the recovery of the pathological synaptic plasticity. This suggested that EGb761 played an important and improving role on learning and memory dysfunction of VD.

Objective: To discuss the diagnosis and treatment of condylar resorption of unknown reason., Methods: The clinical data including the records of history, physical examination, radiography and laboratory of ten patients were studied. Ten patients consisted of 8 females and 2 males (mean age 24.1 years, range 19-31 years) had common clinical features including anterior open bite, posterior occlusal prematurities and Class II malocclusion. Images demonstrated a small and short condyle with abnormal shape, usually accompanied by the developmental insufficiency of the ramus and condyle. The condylar, even the ramus, showed the black marrow signal on MRI images. Four patients were treated by condylar reconstruction with costochondral graft after condylectomy under the supervision of endoscope. Two patients were treated by reduction and repair of disk. Four patients didn't accept any treatment., Results: Four patients treated by condylar reconstruction with costochondral graft showed structures with the size and shape morphologically similar to normal joint and achieved a stable occlusion after follow-up of 6-18 months. Two patients treated by reduction and repair of disk didn't show significant change of open bite and remodeling of condyle after follow-up of 4-6 months., Conclusion: The diagnosis of condylar resorption of unknown reason can be achieved based on the physical examination and radiographical images. The condylar reconstruction with costochondral graft after condylectomy is feasible.

Objective: To discuss the mechanism of mice acute hepatic injury caused by tripterygium glycosides tablets with different dose at different time., Method: Mice were given Tripterygium Glycosides respectively with the dosage of 10 times, 20 times, 30 times of clinical dose to observe the change of mice acute hepatic injury with different does; then, the acute hepatic injury mice were duplicated with 20 times clinical dosage and mice serum ALT were detected at 9, 18, 27, 36 h to observe the change of mice acute hepatic injury at different time. The activity of SOD, GSH-Px in serum and the level of LPO in liver homogenate wevedetected to discuss the mechanism of mice acute hepatic injury caused by Tripterygium Glycosides tablets; and liver tissue pathology was observed., Result: The acute hepatic injury was obvious with 20 times adult dosage in 18 hours and the acute hepatic injury mice death rate was low., Conclusion: Tripterygium Glycosides tablets can cause acute hepatic injury to mice and its mechanism is related to Lipid peroxidation reaction.